我们感谢德意志民主共和国（DFG），KFO 247，为我们的研究提供资金。

实验程序按照德国"动物福利法"（2014年最后修订）和欧洲条例（2010/63 / EU）进行。实验由当地动物福利机构（LaGeSo，Berlin）批准，符合当地部门和国际准则。 注意:在所提出的方法中，使用两个模型的电极从连接主运动皮质（M1）与底丘核（STN）和黑质基质（SNr）连接的超直接皮质基底神经节途径记录。对于使用M1定制低阻抗Ag / AgCl电极的硬膜外皮质电图（ECoG）记录。来自STN和SNr的记录用市售的高阻钨电极进行。 硬膜外Ag / AgCl硬膜外电极的构建<ol><li>拿一个约5厘米长的99.99％纯银线条，带直径r为200μm，如有必要，可以去除任何涂层。 </li><li>将电极尖端向下插入打火机或蜡烛的火焰中，直到尖端开始熔化。等到尖端是球形，直径约为1毫米。将预制的电极从球形尖端的开始到导线端部的总长度为15mm。 </li><li>焊接精密连接器到线端，适合所用的电生理记录系统。用导电银漆覆盖焊丝端到连接器的焊点。这有助于电导率，并产生更好的信号质量。 </li><li>导电清漆干燥后，用3 mm至1 mm的热收缩管覆盖焊点。仔细使用制表师的锤子将球形尖端压平至厚度的一半。 </li><li>戴上检查手套，取100％乙醇的无绒清洁布清除任何污垢和油脂。 </li><li>把电极放入15 mL离心管或细胞培养管，并用家用氯漂白剂（注意:每100g溶剂含有2.8克次氯酸钠），直到球形尖端完全被覆盖。 注意:氯漂白剂有腐蚀性;始终遵循制造商的安全说明。 </li><li> 23分钟后取出电极，用蒸馏水漱口。氯化银层的成功应用表现为均匀的紫色变化。 </li><li>在空气中干燥完全干燥后，用细镊子取电极。使用精美的画笔，应用液体电气绝缘。直接从电极尖端的电线上开始，并覆盖所有的热收缩管。让绝缘干燥至少2小时。 </li><li>为了进行质量控制，请用万用表检查电导率。如果可用，使用适当的阻抗表在1 kHz下执行阻抗测试，同时进行电极和测试将探针放在含有0.9％NaCl的H 2 O溶液中而不相互接触。 1 kHz时的阻抗值应为约8kΩ。 </li></ol> 2.将电极固定在立体定位架上注意:要同时记录MUA和LFP，请使用阻抗为1.5MΩ的钨微丝电极。如果记录的焦点在单个单元的高质量记录上，请选择具有较高阻抗（&gt; 5MΩ）的微线电极。如果研究的目的仅针对LFP，则具有较低阻抗的电极是可以接受的。对于经常需要进行背侧立体定位调整的小型结构，可以使用具有合适的背侧尖端分离（在这种情况下为250μm）的成对电极。此外，如果需要，这提供了更局部的参考电极的优点。始终从最下面的电极和a测量立体定位坐标重新计算参考伯尔贾。 <ol><li>拿一个带有丙烯酸块和夹具的标准立体定位电极支架，并将其牢牢放在手术显微镜视野的平坦表面上。 </li><li>使用细镊子将第一对电极松散地固定在带有胶带（3毫米×8毫米）的支架的丙烯酸块上。电极应突出丙烯酸块约12毫米。 </li><li>仔细地将第二双极电极固定在第一电极的旁边。为了瞄准超直接路径的结构，距离应为2mm（ 图1 ）。对于大多数标准的立体定位电极支架，这是相邻的凹槽。使用卡尺来验证。可以以相同的方式接近不同的网络。为此，可以将丙烯酸嵌段转变到一定程度。 </li><li>通过将第二对电极小心地滑动到最接近顶端约200的位置来调整第二对电极与第一电极相比凹陷（ 图1 ）。在微观视觉下进行。为此，使用30 G插管（外径300μm）来更好地估计距离。 </li><li>按下胶带，然后用夹具的金属夹固定。 </li></ol>手术<ol><li>对于电生理记录，使用聚氨酯（小心）进行麻醉。 注意:氨基甲酸是有毒和致癌的，因此始终遵守物质制造商提供的安全规定和数据表。 </li><li>在0.9％的NaCl食盐水溶液中制备200 mg / mL聚氨酯溶液。 </li><li>腹膜内（IP）总共施用1.3g / kg体重尿布。根据大鼠应变，在注射之间以15分钟间隔将剂量分成两个剂量可能是合理的，以便增强麻醉的安全性。 </li><li>使用p检查麻醉深度戒烟反射和其他合适的反射。麻醉不足以进行手术，注射0.15 g / kg体重的尿烷IP，再等15分钟。 </li><li>应用眼膏防止角膜脱水。 </li><li>在麻醉期间不断监测呼吸频率和踏板撤退反射。使用温度控制的小型动物加热垫，以确保在整个手术过程中保持生理体温。在开始电生理记录之前，更换为非电气替代品（ 如醋酸钠头垫）。 </li><li>剃毛与头部的背侧，以实现一个干净的手术领域。用适当的手术消毒剂在切口部位周围消毒。将动物固定在立体定向框架中。 </li><li>用手术刀在矢状方向进行2厘米长的头皮切口。使用手术刀稍微刮掉头骨腱膜并消毒颅骨。使用公司浸泡在3％H 2 O 2中的豆芽以除去任何剩余的组织。 </li><li>如有必要，请使用电动机或电热器控制出血。停止颅骨和皮下注射出血，如果出血不能在1-2分钟后自发停止，并阻止颅骨的视线。 </li><li>调整门牙杆，直到头部位于平坦的颅骨位置，这意味着作为立体定位参考点的母体和lambda位于同一平面。这是实现高手术精度最重要的。使用标准的立体定位大鼠对齐工具，在微观视觉下校准指定的尖端至胭脂，并调整切牙杆，直到工具上的指定点和工具上的lambda同时触及颅骨。 注意:从一侧的视图与来自另一侧的聚焦光可能有助于确定这种情况。或者，取一个具有精细插管的立体定位支架，并测量Bregma a的背向坐标ndλ在微观视觉下。调整门牙杆，直到Bregma和lambda的背侧坐标相同。 </li><li>使用具有插管的立体定位夹具，校准到母体，然后计算颅骨上所有钻孔的位置。使用立体定位支架，通过仔细抓颅骨或使用手术颜色标记来标记待钻孔的位置。这个坐标取决于目标，参考表格的坐标给出了表1中的超直接路径，包括小脑参考电极的建议坐标。 </li><li>使用微型钻头仔细钻孔。对于STN和SNr，钻一个公共孔（大小为2 mm x 3 mm）。所有其他钻孔应具有约1mm的直径。 </li><li>取两个细管（至少27 G），并弯曲其尖端以形成钩形，使用硬表面或镊子。使用这些来删除任何debris从钻孔中，并在普通STN / SNr孔中仔细切割并去除硬脑膜。 </li><li>用生理盐水冲洗钻孔。每15分钟应用一滴生理盐水给钻孔，以防止脑和硬脑膜变干。 </li><li>用微型钻头和匹配的不锈钢微型螺钉（ 如 M 1.2 mm x 2 mm螺丝），在小脑上方的参考硬膜外电极的钻孔之间钻一个孔并拧入微螺钉，对于M1硬膜外电极</li><li>将自建的Ag / AgCl硬膜外电极滑入参考电极和M1电极的钻孔中。用精细的镊子引导电极尖端，并将其直接滑入颅骨下方进入钻孔。 </li><li>用双组分牙科丙烯酸修复所有硬膜外电极。确保不要覆盖边缘点，也不影响常见的STN / SNr孔。 </li><li>用钨微丝插入准备好的支架进入立体定位框架。 </li><li>校准旨在瞄准STN的大多数腹侧电极到正前方。调整到公共STN / SNr孔以上的计算位置，并在微观视觉下将电极降低到大脑。确保钨微丝电极顺利进入大脑。 </li></ol> 4.电生理测绘与录音注意:对于此步骤，需要具有能够进行在线过滤和在线穗分选的记录软件的法拉第笼和多通道电生理记录系统。优选使用与位于动物头部附近的前置放大器一起工作的系统，以将电噪声和伪像保持在绝对最小值。除了钨微丝电极之外，还需要至少一个硬膜外和一个参比电极来执行超直接通路的记录。建议插入硬膜外和参考e电极成对，没有它们相互接触，这有助于在发生故障的情况下，并允许在数据分析中进行不同类型的参考。 <ol><li>将移动法拉第笼放在立体定位框架上方。如果只有一个固定的法拉第笼可用，请仔细将立体定位框架移动到法拉第笼中，同时确保深脑电极不会下降到脑中，直到立体定位框架处于最终位置。 </li><li>将电极连接到电生理设置的前台。确保参考电极连接到适当的参考通道。 </li><li>设置录音软件:带通滤波器（0.05-8,000 Hz）并放大（增益1,500-2,000x）原始数据信号。使用适当设置的在线LFP和尖峰滤波器（LFP的带通滤波器为0.05-250 Hz，MUA为300-8,000 Hz的带通滤波器）。对于所有过滤器，请使用butterworth型过滤器。 </li><li>为onlin设置峰值阈值（如果适用）e尖峰排序。大多数记录软件允许设置尖峰阈值，其是幅度值，高于该幅度信号由软件标记为尖峰。该阈值可以以数学方式确定为滤波的尖峰信号的平均幅度的因子或标准偏差，或者可以优选地通过目视检查&lt;500ms的数据段来确定，并且在图形中设置为高于信号噪声的线用户界面。 注意:设置尖峰阈值的意图是计数尖峰和排序单位，以提供有关目前记录多少神经元以及它们的尖峰形状的信息。 </li><li>将钨微丝电极慢慢地降低到目标的背面1mm，这是超直接通路的STN。如有必要，请等待信号稳定。 </li><li>对于电生理映射，以100微米的步长将电极推向腹侧。在每一步，评估射击模式，射击r吃的和形状的钉。与图2中给出的典型示例进行比较。通常，密集的核在几个背部步骤中显示出快速连续的尖峰，而富含纤维的结构在随后的腹侧步骤中显示低的燃烧速率和较小的同质刺激活性。 </li><li>对于超直接通路，确保最内侧电极位于STN内。 注意:当检测到Zona incerta腹侧的MUA显着增加时，达到STN。腹部到STN，由于电极已经到达内囊，几乎完全停止。当大多数腹侧电极处于STN中时，钨微电极的结构确保后部，第二电极处于SNr。在STN和SNr同时记录典型的MUA可能需要以小增量进行背心式微调。注意，MUA的频率取决于实际记录的神经元数量和脑水平激活。 </li><li>一旦电极处于所需的结构，设置在线过滤和尖峰分选（ 见图4 ），然后开始记录数据。可以在LFP记录中识别的不同皮层同步状态的典型示例如图3所示。 </li></ol> 5.实验结束<ol><li>录制完成后，将电极慢慢地从脑中提起，并立即用生理盐水冲洗。电极可以在彻底冲洗和目视检查后重新使用。放电弯曲电极。 </li><li>通过IP注射过量的尿烷（2.5g / kg体重）来安乐死动物。 注:聚氨酯只能用于最终程序。 </li><li>如果需要对电极位置进行组织学验证或其他组织学染色手术，请从头骨中取出大脑并进行处理组织适当。 注意:根据预期的染色方法，可能需要经心灌注。对于电极位置的验尸验证，在大多数情况下，标准Nissl染色足以在例如冠状脑切片中观察电极轨迹。促进组织学目标验证的其它方法包括通过在电极插入16,17之前通过记录电极施加电流或应用生物相容性染料，将电诱导的损伤用于脑组织。 </li></ol>

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(56), (2011).</li><li>Mallet, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Disrupted+dopamine+transmission+and+the+emergence+of+exaggerated+beta+oscillations+in+subthalamic+nucleus+and+cerebral+cortex.">Disrupted dopamine transmission and the emergence of exaggerated beta oscillations in subthalamic nucleus and cerebral cortex.</a> J Neurosci. 28 (18), 4795-4806 (2008).</li><li>Steriade, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Corticothalamic+resonance,+states+of+vigilance+and+mentation.">Corticothalamic resonance, states of vigilance and mentation.</a> Neuroscience. 101 (2), 243-276 (2000).</li><li>Maesawa, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-term+stimulation+of+the+subthalamic+nucleus+in+hemiparkinsonian+rats:+neuroprotection+of+dopaminergic+neurons.">Long-term stimulation of the subthalamic nucleus in hemiparkinsonian rats: neuroprotection of dopaminergic neurons.</a> J Neurosurg. 100 (4), 679-687 (2004).</li><li>Paxinos, G., Watson, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The Rat Brain in Stereotaxic Coordinates. , (1998).</li><li>Oliveira, L. M. O., Dimitrov, D., Nicolelis, M. A. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Methods for Neural Ensemble Recordings Frontiers in Neuroscience. , (2008).</li><li>Torres, E. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increased+efficacy+of+the+6-hydroxydopamine+lesion+of+the+median+forebrain+bundle+in+small+rats,+by+modification+of+the+stereotaxic+coordinates.">Increased efficacy of the 6-hydroxydopamine lesion of the median forebrain bundle in small rats, by modification of the stereotaxic coordinates.</a> J Neurosci Methods. 200 (1), 29-35 (2011).</li><li>Hadar, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rats+overexpressing+the+dopamine+transporter+display+behavioral+and+neurobiological+abnormalities+with+relevance+to+repetitive+disorders.">Rats overexpressing the dopamine transporter display behavioral and neurobiological abnormalities with relevance to repetitive disorders.</a> Sci Rep. 6, 39145 (2016).</li><li>Parr-Brownlie, L. C., Poloskey, S. L., Bergstrom, D. A., Walters, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Parafascicular+thalamic+nucleus+activity+in+a+rat+model+of+Parkinson's+disease.">Parafascicular thalamic nucleus activity in a rat model of Parkinson's disease.</a> Exp Neurol. 217 (2), 269-281 (2009).</li><li>Steriade, M., Nunez, A., Amzica, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+slow+(&lt;+1+Hz)+oscillation+of+neocortical+neurons+in+vivo:+depolarizing+and+hyperpolarizing+components.">A novel slow (&lt; 1 Hz) oscillation of neocortical neurons in vivo: depolarizing and hyperpolarizing components.</a> J Neurosci. 13 (8), 3252-3265 (1993).</li><li>Maggi, C. A., Meli, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Suitability+of+urethane+anesthesia+for+physiopharmacological+investigations+in+various+systems.+Part+1:+General+considerations.">Suitability of urethane anesthesia for physiopharmacological investigations in various systems. Part 1: General considerations.</a> Experientia. 42 (2), 109-114 (1986).</li><li>Goldberg, J. A., Kats, S. S., Jaeger, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Globus+pallidus+discharge+is+coincident+with+striatal+activity+during+global+slow+wave+activity+in+the+rat.">Globus pallidus discharge is coincident with striatal activity during global slow wave activity in the rat.</a> J Neurosci. 23 (31), 10058-10063 (2003).</li><li>Karain, B., Xu, D., Bellone, J. A., Hartman, R. E., Shi, W. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rat+globus+pallidus+neurons:+functional+classification+and+effects+of+dopamine+depletion.">Rat globus pallidus neurons: functional classification and effects of dopamine depletion.</a> Synapse. 69 (1), 41-51 (2015).</li><li>Paasonen, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+seven+different+anesthesia+protocols+for+nicotine+pharmacologic+magnetic+resonance+imaging+in+rat.">Comparison of seven different anesthesia protocols for nicotine pharmacologic magnetic resonance imaging in rat.</a> Eur Neuropsychopharmacol. 26 (3), 518-531 (2016).</li><li>Mahmud, M., Vassanelli, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Processing+and+Analysis+of+Multichannel+Extracellular+Neuronal+Signals:+State-of-the-Art+and+Challenges.">Processing and Analysis of Multichannel Extracellular Neuronal Signals: State-of-the-Art and Challenges.</a> Front Neurosci. 10, 248 (2016).</li><li>Hadar, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Altered+neural+oscillations+and+elevated+dopamine+levels+in+the+reward+pathway+during+alcohol+relapse.">Altered neural oscillations and elevated dopamine levels in the reward pathway during alcohol relapse.</a> Behav Brain Res. 316, 131-135 (2017).</li><li>Voget, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Altered+local+field+potential+activity+and+serotonergic+neurotransmission+are+further+characteristics+of+the+Flinders+sensitive+line+rat+model+of+depression.">Altered local field potential activity and serotonergic neurotransmission are further characteristics of the Flinders sensitive line rat model of depression.</a> Behav Brain Res. 291, 299-305 (2015).</li></ol>

作者没有什么可以披露的。

在本研究中，该方法证明了如何使用连接M1与啮齿动物的STN和SNr的超直接皮层 - 基底神经节途径的实例同时从给定网络的多个位点记录细胞外电生理信号。 记录小皮质结构如STN的关键步骤是将记录电极精确地引导到靶中。在所提出的方法中，注意两个关键步骤确保了目标的高准确性。在将电极引入大脑之前，在立体定向装置中制备动物时，绝对必须确保将头骨带入"平头颅骨"?...

最近关于不同神经精神障碍如帕金森病（PD）和精神分裂症的累积证据强烈地表明，他们的病理生理学是基于经常涉及皮层和皮质下结构1,2,3的延长的神经元回路的关键功能障碍。根据这一理论，疾病的临床表现是由于细胞网络的信息处理能力受损，而不是单个细胞或特定神经元素1,2,3的结果 。为了增强对这种复杂的神经精神疾病组的了解，并找到新的治疗方案，必须非常详细地描述人类患者和动物模型中无序网络的神经元动力学。优秀研究生物科目大规模网络的方法是细胞外电位的多位点电生理记录4 。使用这种方法，可以同时评估局部场电位（LFP），其主要表示由突触前电位5产生的兴奋性和抑制性突触后电流和多单位活性（MUA）的时间总和。细胞外电位的记录与研究网络的其他方法（ 例如功能性磁共振成像和钙成像）相比具有几个优点，因为它提供了更高的时间和空间分辨率，并且因为它不依赖于遗传工程生物体5 。然而，细胞外体内记录的使用是有限的，因为它是不能普遍应用的侵入性技术。 体内电生理记录可以在清醒和麻醉的动物中执行排序6 。这两种方法都伴随着具体的利弊。清醒动物的研究允许在执行定义的行为任务期间记录大脑信号，但容易发生运动相关和其他工件7,8 。另一方面，麻醉动物的记录提供了在高度定义的皮层同步状态下用最少的人造物评估LFP和MUA的机会，但是结果在一定程度上也与清醒对象9,10,11中可以发现的有所不同。 近年来，已经证明，LFP的抽样对于描绘网络活动的病理变化特别有用。其中一个突出的例子是人类患者PD的病理生理学研究和动物模型，其中可以显示皮质基底节神经节循环中增强的β振荡与帕金森症运动症状12,13相关 。作为这一研究的结果，目前正在研究β振荡是否可以用作闭环深层脑刺激的在线反馈生物标志物14,15 。 在本研究中，提供了用氨基甲酸酯麻醉的大鼠中LFP和MUA的急性多位点体内电生理记录的详细描述。证明如何通过电生理学使用标准和定制的电极以及如何构建这些电极来完整的网络，例如超直接皮层 - 基底神经节途径。特别强调如何通过co实现基底节神经核的精确靶向将立体定向手术与MUAs的在线注册相结合。

<table>
	<tbody>
		<tr>
			<td>Ag/AgCl custom epidural electrodes</td>
			<td>Goodfellow GmbH 
			D-61213 Bad Nauheim, Germany 
			info@goodfellow.com</td>
			<td>Product-ID AG005127 for 99.99% silver wire</td>
			<td>Ag/AgCl electrodes will allow for better signal quality, but may only be used in acute experiments. Possible replacement: Stainless steel electrodes</td>
		</tr>
		<tr>
			<td>Stereotaxic holder with acrylic block</td>
			<td>David Kopf Instruments, 
			7324 Elmo Street, Tujunga, CA 91042, USA</td>
			<td>Product ID Model 1770 Standard Electrode Holder</td>
			<td>Make sure the acrylic block has recesses which suit the electrode setup for the desired target. Acrylic blocks can easily be modified with a file to obtain the desired configuration. Possible replacement: Self-constructed electrode holders</td>
		</tr>
		<tr>
			<td>Tungsten microwire electrodes 1.5 M&#937; impedance</td>
			<td>Microprobes.com 
			18247-D Flower Hill Way&#160; Gaithersburg, Maryland, 20879 USA</td>
			<td>Product-ID WE3ST31.5A5-250um</td>
			<td>The 1.5 M&#937; is necessary to record MUA and LFP at the same time. Possible replacement: Microelectrodes of different materials can be used. The electrodes have to be straight, robust and as thin as possible.</td>
		</tr>
		<tr>
			<td>Rat alignment tool</td>
			<td>David Kopf Instruments, 
			7324 Elmo Street, Tujunga, CA 91042, USA</td>
			<td>Product ID Model 944 Rat Alignment Tool</td>
			<td>Allows the exact orientation of the brain to match stereotaxic atlases. Possible replacement: Stereotaxic holder with a cannula</td>
		</tr>
		<tr>
			<td>Two-component dental acrylic</td>
			<td>Associated Dental Products Ltd. 
			Kemdent Works, Purton, Swindon 
			Wiltshire, SN5 4HT, United Kingdom</td>
			<td>Simplex Rapid Powder Clear 225g, Product code: ACR803; Simplex Rapid Liquid 150ml, Product code: ACR920</td>
			<td>Depending in the electrodes used, superglue might be an easy alternative, if the electrodes are small and lightweight. Possible replacement: Superglue (Cyanacrylate-based)</td>
		</tr>
		<tr>
			<td>Faraday cage</td>
			<td>Self-construction</td>
			<td></td>
			<td>A proper Faraday cage will be the best protection from electromagnetic artifacts, but everything which can be formed into a box shape or applied to a frame and is made of conductive material may help. Possible replacement: Aluminum foil or copper mesh</td>
		</tr>
		<tr>
			<td>Electrophysiological setup with recording software and online spike-sorting capabilities</td>
			<td>OmniPlex&#174; Neural Data Acquisition System 
			Plexon Inc 
			6500 Greenville Avenue, Suite 700 
			Dallas, Texas 75206 
			USA</td>
			<td></td>
			<td>Offline sorting software is a potential alternative, multiple scripts and softwares can be found for free in the open source community.</td>
		</tr>
	</tbody>
</table>

acute in vivo electrophysiological recordings of local field potentials and multi-unit activity from the hyperdirect pathway in anesthetized rats

融合证据表明，许多神经精神疾病应被理解为大规模神经元网络的紊乱。为了更好地了解这些疾病的病理生理基础，有必要精确地表征电路的不同神经元部分之间的信息处理在哪种方式的干扰。使用细胞外体内电生理记录，可以精确地描绘神经元网络内的神经元活动。该方法的应用与替代技术相比具有几个优点， 例如功能磁共振成像和钙成像，因为它允许独特的时间和空间分辨率，并且不依赖于遗传工程生物体。然而，细胞外体内记录的使用是有限的，因为它是不能普遍应用的侵入性技术。在本文中，介绍一种简单易用的方法可以在网络的多个位置同时记录细胞外电位，例如局部场电位和多单元活动。详细说明如何使用立体定向手术和多单位记录的在线分析来实现皮质下核的精确定位。因此，证明了在麻醉动物体内如何研究完整的网络如超直肠皮质基底神经节环。

使用本文使用的记录电极，可以从STN和SNr从初级运动皮层，下丘脑核和黑质分布样本LFPs和网状细胞。最初，LFP和多单元活动一起记录在宽带信号中。此后，LFP和MUA被带通滤波器分离（LFP为0.05-250Hz，MUA为300-4,000Hz）。 为了正确定位皮质下核，特别是小结构如STN，有利的是将计划的立体坐标与在线记录的MUA信号对齐。对于可以记?...

Watch this Scientific Journal Video about Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats at JoVE.com

Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats

急性<em&gt;体内</em&gt;麻醉大鼠超导途径局部场电位和多单位活动的电生理记录

在本研究中，介绍了如何在尿烷麻醉下从超直接途径进行多位点体内电生理记录的方法。

Converging evidence shows that many neuropsychiatric diseases should be understood as disorders of large-scale neuronal networks. To better understand the ...

acute-vivo-electrophysiological-recordings-local-field-potentials

Research

JoVE Journal

Neuroscience

Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats

15.5K 次观看.  Charité University Medicine Berlin. 在本研究中，介绍了如何在尿烷麻醉下从超直接途径进行多位点体内电生理记录的方法。 

视频: Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats

Electrophysiological Recordings from the Giant Fiber Pathway of D. melanogaster

When startled adult D. melanogaster react by jumping into the air and flying away. In many invertebrate species, including D. melanogaster, the "escape" (or "startle") response during the adult stage is mediated by the multi-component neuronal circuit called the Giant Fiber System (GFS). The comparative large size of the neurons, their distinctive morphology and simple connectivity make the GFS an attractive model system for studying neuronal circuitry. The GFS pathway is composed of two bilaterally symmetrical Giant Fiber (GF) interneurons whose axons descend from the brain along the midline into the thoracic ganglion via the cervical connective. In the mesothoracic neuromere (T2) of the ventral ganglia the GFs form electro-chemical synapses with 1) the large medial dendrite of the ipsilateral motorneuron (TTMn) which drives the tergotrochanteral muscle (TTM), the main extensor for the mesothoracic femur/leg, and 2) the contralateral peripherally synapsing interneuron (PSI) which in turn forms chemical (cholinergic) synapses with the motorneurons (DLMns) of the dorsal longitudinal muscles (DLMs), the wing depressors. The neuronal pathway(s) to the dorsovental muscles (DVMs), the wing elevators, has not yet been worked out (the DLMs and DVMs are known jointly as indirect flight muscles - they are not attached directly to the wings, but rather move the wings indirectly by distorting the nearby thoracic cuticle) (King and Wyman, 1980; Allen et al., 2006). The di-synaptic activation of the DLMs (via PSI) causes a small but important delay in the timing of the contraction of these muscles relative to the monosynaptic activation of TTM (~0.5 ms) allowing the TTMs to first extend the femur and propel the fly off the ground. The TTMs simultaneously stretch-activate the DLMs which in turn mutually stretch-activate the DVMs for the duration of the flight. The GF pathway can be activated either indirectly by applying a sensory (e.g."air-puff" or "lights-off") stimulus, or directly by a supra-threshold electrical stimulus to the brain (described here). In both cases, an action potential reaches the TTMs and DLMs solely via the GFs, PSIs, and TTM/DLM motoneurons, although the TTMns and DLMns do have other, as yet unidentified, sensory inputs. Measuring "latency response" (the time between the stimulation and muscle depolarization) and the "following to high frequency stimulation" (the number of successful responses to a certain number of high frequency stimuli) provides a way to reproducibly and quantitatively assess the functional status of the GFS components, including both central synapses (GF-TTMn, GF-PSI, PSI-DLMn) and the chemical (glutamatergic) neuromuscular junctions (TTMn-TTM and DLMn-DLM). It has been used to identify genes involved in central synapse formation and to assess CNS function.

Electrophysiological Recordings from the Giant Fiber Pathway of D. melanogaster

The Giant Fiber System is a simple neuronal circuit of adult Drosophila melanogaster containing the largest neurons in the fly. We describe the protocol for monitoring synaptic transmission through this pathway by recording post synaptic potentials in dorsal longitudinal (DLM) and tergotrochanteral (TTM) muscles following direct stimulation of the Giant Fiber interneurons.

从巨纤维通路的电生理记录 D。果蝇</em

When startled adult D. melanogaster react by jumping into the air and flying away. In many invertebrate species, including D. melanogaster, the "escape" ...

科研

神经科学

Identification of Olfactory Volatiles using Gas Chromatography-Multi-unit Recordings (GCMR) in the Insect Antennal Lobe

All organisms inhabit a world full of sensory stimuli that determine their behavioral and physiological response to their environment. Olfaction is especially important in insects, which use their olfactory systems to respond to, and discriminate amongst, complex odor stimuli. These odors elicit behaviors that mediate processes such as reproduction and habitat selection1-3. Additionally, chemical sensing by insects mediates behaviors that are highly significant for agriculture and human health, including pollination4-6, herbivory of food crops7, and transmission of disease8,9. Identification of olfactory signals and their role in insect behavior is thus important for understanding both ecological processes and human food resources and well-being.
	To date, the identification of volatiles that drive insect behavior has been difficult and often tedious. Current techniques include gas chromatography-coupled electroantennogram recording (GC-EAG), and gas chromatography-coupled single sensillum recordings (GC-SSR)10-12. These techniques proved to be vital in the identification of bioactive compounds. We have developed a method that uses gas chromatography coupled to multi-channel electrophysiological recordings (termed 'GCMR') from neurons in the antennal lobe (AL; the insect's primary olfactory center)13,14. This state-of-the-art technique allows us to probe how odor information is represented in the insect brain. Moreover, because neural responses to odors at this level of olfactory processing are highly sensitive owing to the degree of convergence of the antenna's receptor neurons into AL neurons, AL recordings will allow the detection of active constituents of natural odors efficiently and with high sensitivity. Here we describe GCMR and give an example of its use.
	Several general steps are involved in the detection of bioactive volatiles and insect response. Volatiles first need to be collected from sources of interest (in this example we use flowers from the genus Mimulus (Phyrmaceae)) and characterized as needed using standard GC-MS techniques14-16. Insects are prepared for study using minimal dissection, after which a recording electrode is inserted into the antennal lobe and multi-channel neural recording begins. Post-processing of the neural data then reveals which particular odorants cause significant neural responses by the insect nervous system.
	Although the example we present here is specific to pollination studies, GCMR can be expanded to a wide range of study organisms and volatile sources. For instance, this method can be used in the identification of odorants attracting or repelling vector insects and crop pests. Moreover, GCMR can also be used to identify attractants for beneficial insects, such as pollinators. The technique may be expanded to non-insect subjects as well.

Identification of Olfactory Volatiles using Gas Chromatography-Multi-unit Recordings GCMR in the Insect Antennal Lobe

Olfactory cues mediate many different behaviors in insects, and are often complex mixtures comprised of tens to hundreds of volatile compounds. Using gas chromatography with multi-channel recording in the insect antennal lobe, we describe a method for the identification of bioactive compounds.

用气相色谱 - 多单元的录音（GCMR）昆虫触角叶嗅觉挥发物的鉴定

All  organisms inhabit a world full of sensory stimuli that determine their  behavioral and physiological response to their environment. Olfaction is ...

Dual Electrophysiological Recordings of Synaptically-evoked Astroglial and Neuronal Responses in Acute Hippocampal Slices

Astrocytes form together with neurons tripartite synapses, where they integrate and modulate neuronal activity. Indeed, astrocytes sense neuronal inputs through activation of their ion channels and neurotransmitter receptors, and process information in part through activity-dependent release of gliotransmitters. Furthermore, astrocytes constitute the main uptake system for glutamate, contribute to potassium spatial buffering, as well as to GABA clearance. These cells therefore constantly monitor synaptic activity, and are thereby sensitive indicators for alterations in synaptically-released glutamate, GABA and extracellular potassium levels. Additionally, alterations in astroglial uptake activity or buffering capacity can have severe effects on neuronal functions, and might be overlooked when characterizing physiopathological situations or knockout mice. Dual recording of neuronal and astroglial activities is therefore an important method to study alterations in synaptic strength associated to concomitant changes in astroglial uptake and buffering capacities. Here we describe how to prepare hippocampal slices, how to identify stratum radiatum astrocytes, and how to record simultaneously neuronal and astroglial electrophysiological responses. Furthermore, we describe how to isolate pharmacologically the synaptically-evoked astroglial currents.

The preparation of acute brain slices from isolated hippocampi, as well as the simultaneous electrophysiological recordings of astrocytes and neurons in stratum radiatum during stimulation of schaffer collaterals is described. The pharmacological isolation of astroglial potassium and glutamate transporter currents is demonstrated.

双突触诱发的星形胶质细胞和神经元的反应，在急性海马脑片电生理记录

Astrocytes form together with neurons tripartite synapses, where they integrate and modulate neuronal activity. Indeed, astrocytes sense neuronal inputs ...

Multi-unit Recording Methods to Characterize Neural Activity in the Locust (Schistocerca Americana) Olfactory Circuits

Detection and interpretation of olfactory cues are critical for the survival of many organisms. Remarkably, species across phyla have strikingly similar olfactory systems suggesting that the biological approach to chemical sensing has been optimized over evolutionary time1. In the insect olfactory system, odorants are transduced by olfactory receptor neurons (ORN) in the antenna, which convert chemical stimuli into trains of action potentials. Sensory input from the ORNs is then relayed to the antennal lobe (AL; a structure analogous to the vertebrate olfactory bulb). In the AL, neural representations for odors take the form of spatiotemporal firing patterns distributed across ensembles of principal neurons (PNs; also referred to as projection neurons)2,3. The AL output is subsequently processed by Kenyon cells (KCs) in the downstream mushroom body (MB), a structure associated with olfactory memory and learning4,5. Here, we present electrophysiological recording techniques to monitor odor-evoked neural responses in these olfactory circuits.
First, we present a single sensillum recording method to study odor-evoked responses at the level of populations of ORNs6,7. We discuss the use of saline filled sharpened glass pipettes as electrodes to extracellularly monitor ORN responses. Next, we present a method to extracellularly monitor PN responses using a commercial 16-channel electrode3. A similar approach using a custom-made 8-channel twisted wire tetrode is demonstrated for Kenyon cell recordings8. We provide details of our experimental setup and present representative recording traces for each of these techniques.

Multi-unit Recording Methods to Characterize Neural Activity in the Locust Schistocerca Americana Olfactory Circuits

We demonstrate variations of the extracellular multi-unit recording technique to characterize odor-evoked responses in the first three stages of the invertebrate olfactory pathway. These techniques can easily be adapted to examine ensemble activity in other neural systems as well.

多单位记录方法来描述蝗虫的神经活动（<em&gt; Schistocerca美洲</em&gt;）嗅觉电路

Detection and interpretation of olfactory cues are critical for the survival  of many organisms. Remarkably, species across phyla have strikingly similar ...

Electrophysiological and Morphological Characterization of Neuronal Microcircuits in Acute Brain Slices Using Paired Patch-Clamp Recordings

The combination of patch clamp recordings from two (or more) synaptically coupled neurons (paired recordings) in acute brain slice preparations with simultaneous intracellular biocytin filling allows a correlated analysis of their structural and functional properties. With this method it is possible to identify and characterize both pre- and postsynaptic neurons by their morphology and electrophysiological response pattern. Paired recordings allow studying the connectivity patterns between these neurons as well as the properties of both chemical and electrical synaptic transmission. Here, we give a step-by-step description of the procedures required to obtain reliable paired recordings together with an optimal recovery of the neuron morphology. We will describe how pairs of neurons connected via chemical synapses or gap junctions are identified in brain slice preparations. We will outline how neurons are reconstructed to obtain their 3D morphology of the dendritic and axonal domain and how synaptic contacts are identified and localized. We will also discuss the caveats and limitations of the paired recording technique, in particular those associated with dendritic and axonal truncations during the preparation of brain slices because these strongly affect connectivity estimates. However, because of the versatility of the paired recording approach it will remain a valuable tool in characterizing different aspects of synaptic transmission at identified neuronal microcircuits in the brain.

Patch-clamp recordings and simultaneous intracellular biocytin filling of synaptically coupled neurons in acute brain slices allow a correlated analysis of their structural and functional properties. The aim of this protocol is to describe the essential technical steps of electrophysiological recording from neuronal microcircuits and their subsequent morphological analysis.

神经微电路急性脑片电生理和形态特征采用配对膜片钳记录

The combination of patch clamp recordings from two (or more) synaptically coupled neurons (paired recordings) in acute brain slice preparations with ...

Simultaneous Recording of Electroretinography and Visual Evoked Potentials in Anesthetized Rats

The electroretinogram (ERG) and visual evoked potential (VEP) are commonly used to assess the integrity of the visual pathway. The ERG measures the electrical responses of the retina to light stimulation, while the VEP measures the corresponding functional integrity of the visual pathways from the retina to the primary visual cortex following the same light event. The ERG waveform can be broken down into components that reflect responses from different retinal neuronal and glial cell classes. The early components of the VEP waveform represent the integrity of the optic nerve and higher cortical centers. These recordings can be conducted in isolation or together, depending on the application. The methodology described in this paper allows simultaneous assessment of retinal and cortical visual evoked electrophysiology from both eyes and both hemispheres. This is a useful way to more comprehensively assess retinal function and the upstream effects that changes in retinal function can have on visual evoked cortical function.

This protocol describes simultaneous measurement of electroretinogram and visual evoked potentials in anesthetized rats.

电图同时记录和麻醉大鼠视觉诱发电位

The electroretinogram (ERG) and visual evoked potential (VEP) are commonly used to assess the integrity of the visual pathway. The ERG measures the ...

Simultaneous Recordings of Cortical Local Field Potentials, Electrocardiogram, Electromyogram, and Breathing Rhythm from a Freely Moving Rat

Monitoring the physiological dynamics of the brain and peripheral tissues is necessary for addressing a number of questions about how the brain controls body functions and internal organ rhythms when animals are exposed to emotional challenges and changes in their living environments. In general experiments, signals from different organs, such as the brain and the heart, are recorded by independent recording systems that require multiple recording devices and different procedures for processing the data files. This study describes a new method that can simultaneously monitor electrical biosignals, including tens of local field potentials in multiple brain regions, electrocardiograms that represent the cardiac rhythm, electromyograms that represent awake/sleep-related muscle contraction, and breathing signals, in a freely moving rat. The recording configuration of this method is based on a conventional micro-drive array for cortical local field potential recordings in which tens of electrodes are accommodated, and the signals obtained from these electrodes are integrated into a single electrical board mounted on the animal's head. Here, this recording system was improved so that signals from the peripheral organs are also transferred to an electrical interface board. In a single surgery, electrodes are first separately implanted into the appropriate body parts and the target brain areas. The open ends of all of these electrodes are then soldered to individual channels of the electrical board above the animal's head so that all of the signals can be integrated into the single electrical board. Connecting this board to a recording device allows for the collection of all of the signals into a single device, which reduces experimental costs and simplifies data processing, because all data can be handled in the same data file. This technique will aid the understanding of the neurophysiological correlates of the associations between central and peripheral organs.

This study introduces a method for the simultaneous recording of local field potentials in the brain, electrocardiograms, electromyograms, and breathing signals of a freely moving rat. This technique, which reduces experimental costs and simplifies data analysis, will contribute to the understanding of the interactions between the brain and peripheral organs.

自由运动大鼠皮质局部电位、心电图、肌电图和呼吸节律的同步记录

Monitoring the physiological dynamics of the brain and peripheral tissues is necessary for addressing a number of questions about how the brain controls ...

Quantifying Acute Changes in Renal Sympathetic Nerve Activity in Response to Central Nervous System Manipulations in Anesthetized Rats

Renal sympathetic nerve activity (RSNA) and mean arterial pressure are important parameters in cardiovascular and autonomic research; however, there are limited resources directing scientists in the techniques for measuring and analyzing these variables. This protocol describes the methods for measuring RSNA and mean arterial pressure in anesthetized rats. The protocol also includes the approaches for accessing the brain during RSNA recordings for central nervous system (CNS) manipulations. The RSNA recording technique is compatible with pharmacologic, optogenetic, or electrical stimulation of the CNS. The approach is useful when an investigator will measure short-term (min to h) autonomic responses in non-survival experiments to correlate anatomically with CNS nuclei. The approach is not intended to be used to obtain chronic (survival) recordings of RSNA in rats. Discharges in RSNA, averaged rectified RSNA, and mean arterial pressure can be quantified and analyzed further using parametric statistical tests. Methods for obtaining venous access, recording mean arterial pressure telemetrically, and brain fixation for future histological analysis are also described in the article.

Methods for measuring sympathetic and cardiovascular responses to central nervous system (CNS) manipulations are important for advancing neuroscience. This protocol was developed to assist scientists with measuring and quantifying acute changes in renal sympathetic nerve activity (RSNA) in anesthetized rats (non-survival).

麻醉大鼠中枢神经系统操作对肾交感神经活动的急性变化量化

Renal sympathetic nerve activity (RSNA) and mean arterial pressure are important parameters in cardiovascular and autonomic research; however, there are ...

Labeling and Imaging of Amyloid Plaques in Brain Tissue Using the Natural Polyphenol Curcumin

Deposition of amyloid beta protein (A&#946;) in extra- and intracellular spaces is one of the hallmark pathologies of Alzheimer&#39;s disease (AD). Therefore, detection of the presence of A&#946; in AD brain tissue is a valuable tool for developing new treatments to prevent the progression of AD. Several classical amyloid binding dyes, fluorochrome, imaging probes, and A&#946;-specific antibodies have been used to detect A&#946; histochemically in AD brain tissue. Use of these compounds for A&#946; detection is costly and time consuming. However, because of its intense fluorescent activity, high-affinity, and specificity for A&#946;, as well as structural similarities with traditional amyloid binding dyes, curcumin (Cur) is a promising candidate for labeling and imaging of A&#946; plaques in postmortem brain tissue. It is a natural polyphenol from the herb Curcuma longa. In the present study, Cur was used to histochemically label A&#946; plaques from both a genetic mouse model of 5x familial Alzheimer&#39;s disease (5xFAD) and from human AD tissue within a minute. The labeling capability of Cur was compared to conventional amyloid binding dyes, such as thioflavin-S (Thio-S), Congo red (CR), and Fluoro-jade C (FJC), as well as A&#946;-specific antibodies (6E10 and A11). We observed that Cur is the most inexpensive and quickest way to label and image A&#946; plaques when compared to these conventional dyes and is comparable to A&#946;-specific antibodies. In addition, Cur binds with most A&#946; species, such as oligomers and fibrils. Therefore, Cur could be used as the most cost-effective, simple, and quick fluorochrome detection agent for A&#946; plaques.

Curcumin is an ideal fluorophore for labeling and imaging of amyloid beta protein plaques in brain tissue due to its preferential binding to amyloid beta protein as well as its structural similarities with other traditional amyloid binding dyes. It can be used to label and image amyloid beta protein plaques more efficiently and inexpensively than traditional methods.

使用天然多酚姜黄素对脑组织淀粉样斑块进行标记和成像

Deposition of amyloid beta protein (A&#946;) in extra- and intracellular spaces is one of the hallmark pathologies of Alzheimer&#39;s disease (AD). ...

Establishment of Acute Pontine Infarction in Rats by Electrical Stimulation

Pontine infarction is the most common stroke subtype in the posterior circulation, while there lacks a rodent model mimicking pontine infarction. Provided here is a protocol for successfully establishing a rat model of acute pontine infarction. Rats weighing about 250 g are used, and a probe with an insulated sheath is injected into the pons using a stereotaxic apparatus. A lesion is produced by the electrical stimulation with a single pulse. The Longa score, Berderson score, and beam balance test are used to assess neurological deficits. Additionally, the adhesive-removal somatosensory test is used to determine sensorimotor function, and the limb placement test is used to evaluate proprioception. MRI scans are then used to assess the infarction in vivo, and TTC staining is used to confirm the infarction in vitro. Here, a successful infarction is identified that is located in the anterolateral basis of the rostral pons. In conclusion, a new method is described to establish an acute pontine infarction rat model.

Presented here is a protocol for establishing acute pontine infarction in a rat model via electrical stimulation with a single pulse.

电刺激在大鼠中建立急性庞廷梗死

Pontine infarction is the most common stroke subtype in the posterior circulation, while there lacks a rodent model mimicking pontine infarction. Provided ...